Mcf 10a

HeLa (CCL-2), MCF-7 (HTB-22) and, MDA-MB231 (HTB-26) cells were obtained from American Type Culture Collection (ATCC) (Manassas, VA) and maintained in DMEM supplemented with 10% fetal bovine serum, Bio-techne (Minneapolis, MN) and 1% Penicillin-Streptomycin, Thermo Fisher Scientific (Waltham, MA). MCF-10A (ATCC-CRL-10317) cells were a kind gift from Dr. Chambers at the University of Arkansas for Medical Sciences (UAMS) and were maintained in Mammary Epithelial Basal Cell Medium, Promo Cell (Heidelberg, Germany) supplemented with Mammary Epithelial Cell Growth Medium Supplement Pack, Promo Cell (Heildelberg, Germany) in addition to 100μg/mL gentamicin, Sigma-Aldrich (St. Louis, MO) and 0.05μg/mL amphotericin B, Sigma-Aldrich (St. Louis, MO). All cells were grown at 37⁰C in an incubator with 5% CO₂.

Different cell lines (all from American Type Culture Collection ATCC, Manassas, VA, USA) were used to compare the results of primarily isolated cells to established models. The mammary epithelial cell line MCF-10A (ATCC® CRL-10317™) was cultivated in Mammary Epithelial Cell Growth Medium (PromoCell GmbH, Heidelberg, Germany) with the addition of 100 ng/ml cholera toxin (Sigma Aldrich). Furthermore, we used six different breast cancer cell lines. BT-474 (ATCC® HTB-20™, luminal B) were cultivated in Hybri-Care Medium (ATCC), 10% FBS, 1.5 g/l NaHCO₃ (Sigma Aldrich). MCF-7 (ATCC® HTB-22™, HR-positive) were cultivated in DMEM, 10% FBS, 2 mM L-glutamine. MDA-MB-231 (ATCC® HTB-26™, TNBC, mesenchymal-like) were cultivated in DMEM, 10% FBS, 2 mM L-glutamine. SKBR3 (ATCC® HTB-30™, HER2-positive) were cultivated in McCoy’s 5a Modified Medium (Thermo Fisher Scientific Inc.), 10% FBS. MDA-MB-468 (ATCC® HTB-132™, TNBC, basal-like) were cultivated in DMEM/F12 Medium (Biochrom), 10% FBS. T-47D (ATCC® HTB-133™, Luminal A) were cultivated in RPMI-1640 (Thermo Fisher Scientific Inc.), 10% FBS.